1 Introduction

The diamondback moth (Plutella xylostella L.) is an important pest in the family Plutellidae of the order Lepidoptera. It is widely distributed around the world and is one of the most destructive pests of cruciferous crops globally. Recent studies show that this pest has spread to 128 countries and regions, making it the most widely distributed lepidopteran pest in the world (Furlong et al., 2021). Its special biological characteristics make it highly harmful. Under suitable temperatures (Around 25°C), it can complete one generation in only 16~19 days (Huaripata and Sánchez, 2019). Female moths have strong reproductive ability, laying an average of 200 to 300 eggs (Ram et al., 2016). It is also very adaptable and can survive and reproduce in a wide range of areas, from tropical to temperate regions (from 60°N to 40°S) (Philips et al., 2014). These abilities make the diamondback moth very hard to control.

In China, the number of diamondback moth generations each year depends on the region. In northern areas, there are usually 3 to 6 generations. Because greenhouses are more common now, the moth can survive farther north—about 2 to 3 degrees higher in latitude than before (Zhen, 2011). In the south, the moth can have 10 to 15 generations a year. In tropical places like Hainan, it can breed all year round (Yang et al., 2015). The larvae feed on leaves and make "window-like" holes. The moth is also very resistant to pesticides, especially to pyrethroid pesticides, which are several times higher than normal levels of resistance (Soleymanzade et al., 2019). Controlling this pest is expensive. Worldwide, it costs more than 4 billion USD every year (Javier et al., 2019), which shows how serious the problem is.

Right now, many control methods for the diamondback moth have made good progress. In farming, rotating cruciferous crops with non-host plants helps break the pest’s life cycle and lowers the number of places it can live. Clearing crop leftovers after harvest can also cut down overwintering sources (Shakeel et al., 2017). Physical control has improved too. Pheromone traps, like LED light traps, are used to attract and catch adult moths (Reddy and Guerrero, 2000). In biological control, natural enemies like parasitoid wasps (Cotesia plutellae) and predators (Nabis capsiformis) help reduce moth numbers. Some fungi, such as Beauveria bassiana, are used as bio-pesticides to kill the larvae (Nunes et al., 2019; Ramírez-Cerón et al., 2022). For chemical control, newer insecticides like chlorantraniliprole work well-over 90% effective-and are less harmful to helpful insects (Soleymanzade et al., 2019). But it’s important to rotate these chemicals to prevent the moth from becoming resistant.

The goal of this study is to build a reliable lab system to raise diamondback moths. We observed their growth and behavior and tested how different conditions affect them. This helps provide standard moth populations for future research, including resistance tests and the development of greener pest control methods. Our results offer both basic knowledge and technical support for safer vegetable production.

2 Materials and Methods

2.1 Rearing preparation

The diamondback moth (Plutella xylostella) is a typical monophagous pest that mainly feeds on cruciferous plants, especially Chinese cabbage, Chinese kale, and cauliflower (Paudel et al., 2022). Its host plant preference is closely related to glucosinolates and their breakdown products, isothiocyanates. These volatile substances strongly attract female moths to lay eggs and act as chemical signals for host recognition (Hussain et al., 2021).

Store-bought Chinese cabbage was chosen as the host plant because it has strong adaptability, high water content, and is rich in glucosinolates. These traits help the moths lay more eggs and encourage the larvae to feed. Chinese cabbage is also easy to grow and manage in the lab, which helps keep the food quality steady. Recent studies show that using Chinese cabbage can improve larval survival and help them grow faster compared to other plants (Chen et al., 2023).

In the early larval stage, the moth is very sensitive to the smell of glucosinolates (Badenes-Pérez et al., 2013; Badenes-Pérez et al., 2019). The young larvae use the scent from leaf surfaces to find food. To make sure the newly hatched larvae could feed properly, this experiment used fresh, tender Chinese cabbage seedlings with healthy leaves.

2.2 Experimental methods

This experiment was done indoors under controlled conditions. Adult moths were collected from the field, and Chinese cabbage was used as the food plant. The goal was to observe the full life cycle of the diamondback moth. The experiment included five main steps: (1) Collecting adult moths from the field and separating males and females to start the first generation; (2) Growing Chinese cabbage at the same time to use as the host plant; (3) Setting up a controlled environment with fixed temperature, humidity, and light cycle; (4) Rearing and watching the moth through all life stages-egg, larva, pupa, and adult; (5) Recording how long each stage took, how the insects behaved, and how their numbers changed (Figure 1). This setup was based on recent lab methods for rearing diamondback moths. It helps provide a steady insect supply and creates clear, repeatable data (Xing et al., 2014).

2.3 Rearing conditions and experimental tools

2.3.1 Material preparation

Insect collection: Adult moths were collected from a 4m × 4m Chinese cabbage field using insect nets. Host plant: Chinese cabbage seeds were bought from the market. More than 50 plants were sown each time, with 30 cm spacing between plants and rows. Supplementary feeding: Extra cabbage seedlings were planted when needed to ensure enough food for the larvae.

2.3.2 Rearing equipment

Common insect rearing equipment in the lab was used and adjusted based on current rearing needs. These included: Constant-temperature insect-rearing room (±1°C accuracy); Light shelves with adjustable height (30–40 cm); Humidifier (YC-X100 model); Incandescent lamps (for light control); Rearing cages (for pupae and eggs, covered with fine mesh); Syringes (25 mL, for controlled watering); Insect nets, plastic trays, round seedling pots, tweezers, brushes, spray bottles, etc. Recent studies recommend using stacked mesh cages to save space in artificial rearing setups and adding humidifying and ventilation systems to simulate natural conditions. These measures help improve the moth's reproduction success and development synchronization (Zhu et al., 2021).

2.3.3 Indoor rearing environment settings

Temperature: 24°C-28°C (24°C is optimal); Relative humidity: 60%-80%; Lighting: Natural light plus incandescent lamp assistance; Photoperiod: 14 hours light / 10 hours dark (14:10, L:D); Ventilation: Maintain air flow in closed environments to prevent disease buildup

2.4 Field collection of adult moths and host plant cultivation

Adult diamondback moths (Plutella xylostella) are strongly attracted to the smell of cruciferous plants, especially volatile compounds from glucosinolates. These odors help guide egg-laying behavior (Sun et al., 2009). Based on this, a 4m × 4m cabbage plot was chosen for field collection. Active moths were caught using insect nets around sunset.

After collection, moths were sorted by wing color (males: gray-brown; females: gray-white). Adults were placed in egg-laying cages at a female-to-male ratio of 3:2 to form the initial lab population, helping to avoid inbreeding problems. The collection area was set downwind of the cabbage plot to increase moth gathering and improve collection efficiency. Cabbage, a main host plant of the moth, contains high levels of isothiocyanates, which strongly encourage feeding and egg-laying (Badenes-Pérez et al., 2020). Cabbage cultivation was done in two stages:

Plastic mesh shelter stage: Healthy, pest-free seedlings were planted in round pots with soil depth no more than two-thirds of the pot. When seedlings reached 20 mm with open cotyledons, they were moved to square trays to provide more feeding space.

Constant-temperature room stage: When seedlings grew to 40 mm and cotyledons were fully spread, they were moved into the constant-temperature insect-rearing room for use in pupation and egg laying. The room was kept at about 24°C with 60%-80% humidity and a 14:10 (L:D) light cycle. Irrigation was done with 25 mL syringes to avoid overwatering and root rot. Soil was kept moist but not wet.

2.5 Handling methods for each rearing stage

2.5.1 Egg handling

Adult moths can mate and lay eggs the day after emergence. In the experiment, Chinese cabbage seedlings with eggs were moved into egg-laying cages for hatching. The eggs are long oval in shape, light yellow, smooth, and laid either alone or in small groups of 3~5.

During the hatching process, closely inspect the condition of cabbage seedlings to avoid mold growth and prevent damage to eggs or larvae. Research has shown that fungi such as Beauveria bassiana and Metarhizium anisopliae are the main causes of fungal death, especially in high humidity conditions (Chen et al., 2023). The incubation time is greatly affected by temperature. In this study, eggs need about 6 days to hatch at 25 ° C, which is within the typically reported range of 4 to 8 days.

2.5.2 Larval rearing

The larvae go through four instars. The 1st and 2nd instar larvae are very small and feed on leaf tissue, often hiding under the leaf surface and creating transparent “windows.” At this stage, it is important to monitor where they feed and make sure they have enough food.

The 3rd and 4th instar larvae eat much more and can quickly destroy entire leaves, sometimes killing the whole plant. In the experiment, the "lure transfer" method was used to prepare fresh cabbage seedlings in advance, and the scent of mustard oil was used to induce active migration of larvae. Alternatively, a brush could be used to gently transfer the insect body. At this stage, it is necessary to supplement food sources and reduce density in a timely manner to prevent stress behaviors such as biting and self harm caused by competition, and to improve survival rate and developmental synchronicity (Zhu et al., 2021).

2.5.3 Pupa handling

Most larvae pupate by spinning cocoons on the surface of leaves or between stems. The pupae start out green and later turn orange-yellow or gray-brown. They are spindle-shaped, 5–8 mm long, and slightly shiny with a thin white cocoon covering.

In the experiment, pupae were gently picked up with tweezers and placed in petri dishes lined with moist filter paper. They were sorted by color stage to help with synchronized emergence. If there weren’t enough cabbage seedlings, pupae could be stored short-term at low temperature (10°C for 15~20 days), but emergence rate may drop due to low temperatures affecting enzyme activity (Li et al., 2022).

2.5.4 Adult rearing

Adults usually emerge 5~8 days after pupation, mostly around dusk. They are very active and strongly attracted to light. Female moths have strong mating signals. Each female can lay 100–300 eggs and may mate several times in her life.

To keep adults active and extend their egg-laying period, 10% honey water was provided using cotton soaked in the solution, and it was changed daily. To avoid inbreeding problems, wild moths were added to the lab population from time to time to increase genetic diversity.

2.6 Observation indicators and recording methods

The main observation indicators include: egg stage length (time required from egg laying to larval hatching), larval life span (time required from egg hatching to pupation, with attention to age classification), pupal stage length (time required from pupa formation to adult emergence), adult lifespan and egg laying, etc.

The specific recording method is as follows: check the color change and hatching status of the eggs every 12 hours from the day of egg laying, and record the time required until the larvae hatch; after the larvae hatch, observe the larvae's feeding status and growth and development progress once in the morning and evening every day; in the pupal stage, count the number of days from pupation to eclosion, and pay attention to record the color change of the pupa; after the adult eclosion, count the eclosion day as 1 day of adult age, and then observe its mating and egg-laying situation until natural death and record the life span. During the egg-laying stage of adults, collect and count the eggs laid by them every day, and count the total egg-laying amount and egg-laying cycle of a single female in her lifetime.

3 Results and Analysis

3.1 Observation of the life cycle of diamondback moth

Under controlled conditions (temperature 24~28°C, humidity 60%~80%, light/dark cycle 14:10), the diamondback moth takes about 13~20 days to complete one generation. The specific development stages are as follows: (1) Egg stage: about 5 days; (2) Larval stage: 3-11 days, with 4 instars; (3) Pupal stage: 5~8 days; (4) Adult stage: about 3-5 days. Moths may mate on the day they emerge, and start laying eggs 1–2 days after mating (Figure 2).

Compared with recent studies, the development period in this experiment is similar to the reported full life cycle of the diamondback moth at 25°C, which is about 19 days on average (Huaripata and Sánchez, 2019). In addition, different host plants can also affect the development speed. For example, diamondback moths reared on cauliflower had the shortest generation time (average 39.5 days) and the highest number of eggs laid (Huaripata and Sánchez, 2019; Jaleel et al., 2019).

3.2 Morphological characteristics and habits of each life stage

3.2.1 Adult

The adult diamondback moth is 6-7 mm long with a wingspan of 12-15 mm. There are three yellow-white wavy lines on the forewings that form a diamond-shaped mark. Adults are strongly attracted to light and are mainly active at night. During the day, they usually hide on the underside of leaves or in weeds.

In indoor rearing, moths were observed to mate as early as the day after emergence. Mating lasted between 30 and 90 minutes. Female moths can lay 100~300 eggs, and in some cases, more than 500 eggs. Female moths usually emerge earlier than males, which increases the overlap of generations.

The sex ratio is about 5:4. Eggs are mainly laid on the underside of leaves, near the veins or feeding marks. Females show "selective repeated egg-laying" behavior. This matches recent studies suggesting that egg-laying sites are influenced by both the host plant's volatile compounds and surface microstructures (Zhu et al., 2021).

3.2.2 Larva

Newly hatched larvae are brown, and then turn green as they grow. The body length can reach 10~12 mm. First and second instar larvae usually feed inside the leaf tissue, creating semi-transparent “window-like” damage. Third and fourth instars feed on the leaf surface and make holes. When the damage is serious, the leaf may look like a net (Figure 3). When disturbed, larvae show defensive behaviors such as strong wriggling, playing dead, and dropping down on silk threads. In the experiment, it was observed that when larval density was too high or food was lacking, they often bit each other, and sometimes even attacked pupae.

According to Liu et al. (2002), under good temperature control (20°C~28°C), the larval stage of the diamondback moth usually lasts 10~12 days. However, under poor nutrition or crowded conditions, the larval period can extend to more than 15 days, with a higher death rate.

3.2.3 Pupa

Pupation mostly happens on the underside or edge of leaves, where the larvae spin cocoons. Pupae are light green at first, then gradually turn orange-yellow or dark brown. The body length is about 5~8 mm (Figure 4). The pupal shell is thin and semi-transparent, and the internal structures are often visible.

In the experiment, a color-based grouping method was used to sort pupae, so that the emergence period was more concentrated. This made it easier to collect moths at the same time. Guo and Qin (2010) pointed out that temperature and humidity have a strong effect on the emergence rate. At 33°C with high humidity (90% RH), the emergence time was much shorter, and female moths emerged earlier than males.

3.3 Effects of rearing conditions on diamondback moth

3.3.1 Temperature

At 24°C, the hatching rate of diamondback moth eggs was the highest, and larval development was more uniform, which is suitable for continuous rearing. Recent studies show that the optimal temperature for diamondback moth growth is 25±2°C, with an intrinsic growth rate (r) of up to 0.18 day⁻¹. When the temperature goes above 30°C, both survival rate and egg production drop significantly (Jaleel et al., 2019).

2.3.2 Humidity control

When humidity is kept between 60% and 80%, the moths perform well. If humidity is too low, hatching rates decrease; if it is too high, mold infections are more likely to occur. Studies also report that when humidity goes over 85%, larval mortality increases by more than 10% (Guo and Qin, 2010).

3.3.3 Light regulation

Diamondback moths are attracted to light, especially to incandescent lamps. In the experiment, a 14:10 light/dark cycle was used to promote mating and feeding. Campos (2008) also confirmed that longer light periods can increase female egg-laying by about 15% and shorten the egg hatching period.

4 Discussion

In this study, the full life cycle of the diamondback moth (Plutella xylostella) was successfully completed under controlled conditions. The process included collecting moths from the field, growing host plants, and observing each life stage in the lab. The study focused on how temperature, humidity, and light affect the moth’s growth, behavior, and reproduction. When the temperature was set between 24°C~28°C, humidity at 60%~80%, and the light-dark cycle at 14:10, the development from egg to adult took about 13 to 20 days. Each stage had clear physical features that were easy to identify. The moths fed well on Chinese cabbage and adapted well to it. The larvae had a high survival rate, and female moths laid most of their eggs in a short period. The moths were able to keep reproducing continuously indoors. This experiment established a stable lab population of diamondback moths, providing a reliable source for future research on insecticides, behavior, and resistance.

The development of the diamondback moth is very sensitive to temperature and humidity. At 24°C, egg hatch rate was highest, larval development was even, pupal stage was shorter, and adults emerged in a more concentrated period. This helped with collecting insects at the same time. If the temperature goes above 30°C, development speeds up, but the death rate increases and reproduction drops (Jaleel et al., 2019).

Humidity between 60% and 80% supports growth at all life stages. If humidity is too low, egg hatching slows down and larval survival drops. If it is too high, mold problems can happen, which increases the risk of pupae rotting and larvae getting sick (Guo and Qin, 2010). The 14-hour light period helped adults stay active, increased mating, and extended the egg-laying period. Campos (2008) found similar results, and this light cycle is now widely used in standard lab rearing of diamondback moths.

Using Chinese cabbage as the only food source, larvae showed good feeding, and females laid eggs in a short period. Compared with other host plants like cauliflower, radish, and cabbage, survival rate and development speed on Chinese cabbage were in the middle range, but reproduction rate was higher on cauliflower (Liu et al., 2002). In the future, the host plant can be chosen based on the purpose of the study. For example, cauliflower is better for egg-laying behavior studies, and a mix of cruciferous vegetables is better for large-scale rearing (Golizadeh et al., 2009a; Golizadeh et al., 2009b).

This study built a stable and repeatable rearing system for the diamondback moth, which can provide insects at all life stages at the same time. Compared with field collection, lab rearing is easier for controlling conditions and ensures a steady insect supply. This system can be used in many types of research, including pesticide testing, resistance studies, behavior observation, pheromone disruption experiments, and host preference analysis. It also supports the development of new biological control tools. At the same time, this method helps local agricultural extension and supports the connection between lab research and field application.

Acknowledgments

This research was conducted at the Hainan Institute of Tropical Agricultural Resources. I sincerely thank the institute for providing excellent research conditions and a supportive environment. I am especially grateful to my academic advisor, Professor Fang X.J., for his thoughtful guidance and valuable suggestions in topic selection, experimental design, and manuscript preparation.

Conflict of Interest Disclosure

The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest.

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